Recent studies have demonstrated the EBV-specific CTLs are highly effective for prophylaxis of therapy of EBV-lymphoma in immunocompromised patients. However, widespread application of this emerging treatment strategy and extension to other EBV- associated lymphoproliferative diseases (LPDs) will require a reliable and efficient means to generate sufficient quantities of tumor antigen-specific CTLs for effective patient therapy. The primary objective of the proposed studies is to develop and implement a unique process for ex vivo derivation, antigen- specific expansion in response to autologous EBV-transformed lymphoblastoid cell lines or LMP2a-transduced dendritic cells, and retroviral-mediated gene-marking of EBV-specific CTLs in a perfused clinical-scale bioreactor system. The hypothesis that continuous single-pass medium exchange will enable enhanced T- cell production, function, and genetic transduction using flow- through technology in the AastromReplicell TM Cell Production System will be tested. The Phase II program will focus on optimization and full automation of the processes in compliance with GMP regulations. The automated bioreactor system will be tested in clinical trials to assess the safety and efficacy of gene-marked EBV-specific CTLs for therapy of EBV-associated lymphoma and Hodgkin s disease. An automated clinical scale bioreactor system would enable efficient and reliable production of highly functional gene-marked T-cells for antigen-specific immunotherapy of EBV-associated LPDs and other cancers. PROPOSED COMMERCIAL APPLICATION A closed, GMP-compliant, bioreactor system for ex vivo derivation, expansion and gene-marking of therapeutic quantities of EBV-specific CTLs would have immediate commercial value for prevention and treatment of EBV-associated LPDs in immunocompromised patients. The development of an automated process for antigen-specific T-cell expansion will reduce open- process steps and associated costs in equipment and labor, thus enabling widespread application of T-cell-based cancer immunotherapies in a reliable and cost-effective fashion.